Methods: In a parallel group
placebo controlled trial patients were treated with HICS (n=10) or placebo
(n=10) over 26 weeks, with a follow up open label treatment for 26 weeks. Serum samples at baseline, 6 weeks, 26 and 52
weeks were analysed using a multiplex assay for 40 cytokines and growth factors
(Luminex®) and additional individual solid phase
immunoassays for procollagen III N-terminal propeptide
(PIIINP), serum IL-2R, cartilage oligomeric matrix
protein (COMP), TGF-beta1 and von Willebrand factor (vWF). To explore
patterns of change over multiple analytes heat maps were constructed using CIMminer NIH software. Clustering was performed using
correlation, and Pearson coefficient and significance analysis of microarrays
(SAM) correction.

Results: Cluster analysis
defined factors that were increased or decreased from baseline after 26 weeks
treatment with HICS. Results for key
analytes are summarised in Table 1. Consistent with previous preclinical studies
there was evidence for marked upregulation of the hypothalmo-pituitary-adrenal
axis from 6 weeks after HCS treatment and this effect was maintained at 26
weeks. This was evidenced by increase in
alpha-MSH and ACTH in cases treated with HICS.
There were changes in markers of fibroblast biology including changes inbFGF, PIIINP and COMP. Novel findings include consistent increase in
PIIINP associated with improved MRSS suggesting that this may be a marker of
ECM turnover rather than fibrogenesis. Other factors that were frequently reduced,
though not reaching statistical significance, included TIMP2, fractalkine and TGFbeta1 levels.

Conclusion: This study illustrates the feasibility of
conducting relatively short term parallel group placebo controlled trials in
established dcSSc to target skin fibrosis. The benefit of including multiplex
analysis of serum proteins in early phase trials to better understand treatment
mechanisms and disease biology is confirmed.
This study suggests possible mechanisms of action for HICS, including
upregulation of alpha-MSH, that has been shown to be antifibrotic in
preclinical studies, and suggests possible serum markers to be included in
future trials.